Abstract
BackgroundThe larval stage of the model organism Drosophila is frequently used to study host-pathogen interactions. During embryogenesis the cellular arm of the immune response, consisting of macrophage-like cells known as plasmatocytes, is extremely motile and functions to phagocytise pathogens and apoptotic bodies, as well as produce extracellular matrix. The cellular branch of the larval (post-embryonic) innate immune system consists of three cell types—plasmatocytes, crystal cells and lamellocytes—which are involved in the phagocytosis, encapsulation and melanisation of invading pathogens. Post-embryonic haemocyte motility is poorly understood thus further characterisation is required, for the purpose of standardisation.MethodologyIn order to examine post-embryonic haemocyte cytoskeletal dynamics or migration, the most commonly used system is in vitro cell lines. The current study employs an ex vivo system (an adaptation of in vitro cell incubation using primary cells), in which primary larval or pre-pupal haemocytes are isolated for short term analysis, in order to discover various aspects of their behaviour during events requiring cytoskeleton dynamics.SignificanceThe ex vivo method allows for real-time analysis and manipulation of primary post-embryonic haemocytes. This technique was used to characterise, and potentially standardised, larval and pre-pupal haemocyte cytoskeleton dynamics, assayed on different extracellular matrices. Using this method it was determined that, while larval haemocytes are unable to migrate, haemocytes recovered from pre-pupae are capable of migration.
Highlights
Drosophila melanogaster is a robust model organism, with each life stage providing novel aspects for research into the cellular innate immune response
Characterisation of haemocyte motility within the embryo showed that haemocytes can be considered a ‘jack-of-all-trades’ whereby they are implicated in general embryonic development, cellular innate immunity and wound healing, which is similar to an inflammation response [5,6]
The different life stages An analysis of haemocyte behaviour at each larval instar was conducted in order to determine whether there was any difference in their centroid and plasma-membrane morphology, and protrusion activity
Summary
Drosophila melanogaster is a robust model organism, with each life stage providing novel aspects for research into the cellular innate immune response. Drosophila embryos provide an in vivo platform in which the powerful genetic tools of Drosophila can be utilised to manipulate various proteins and pathways, such as Rho-family small GTPases, in order to elucidate the mechanisms controlling cell migration within an organism [3,4]. During embryogenesis the cellular arm of the immune response, consisting of macrophage-like cells known as plasmatocytes, is extremely motile and functions to phagocytise pathogens and apoptotic bodies, as well as produce extracellular matrix. The cellular branch of the larval (post-embryonic) innate immune system consists of three cell types— plasmatocytes, crystal cells and lamellocytes—which are involved in the phagocytosis, encapsulation and melanisation of invading pathogens. Post-embryonic haemocyte motility is poorly understood further characterisation is required, for the purpose of standardisation
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